人工雌核发育大黄鱼(Pseudosciaena crocea)的AFLP分析

采用6对选择性扩增引物(E-AAG/M-CAG、E-AAG/M-CTC、E-ACA/M-CAA;E-ACG/M-CAT、E-AGG/M-CAA、E-AGG/M-CTA)对冷休克法诱导的大黄鱼雌核发育家系G1、G2和对照组C1、C2的鱼苗及其亲本进行了扩增片段长度多态性(AFLP)标记的比较分析。结果表明,6对引物共检出了33条雄亲特有条带(G1中13条,G2中20条)、31条雌亲特有条带(G1中16条,G2中15条)。家系G1的24尾鱼苗均无雄亲特有条带,雌核发育成功率为100%,G2家系中有3尾鱼苗各出现了不同数量的雄亲特有条带,属正常受精个体(12.5%),其雌核发育成功率为87.5%,两个家系平均雌核发育诱导成功率为93.75%。31条雌亲特有条带中有14条在雌核发育后代中出现了分离。对亲子间遗传关系的分析表明了雌核发育个体之间的遗传差异最小,与雌亲的亲缘关系最近。研究表明,雌核发育是促进基因纯合的一个有效途径,AFLP技术是鱼类雌核发育鉴定和遗传分析的有效方法。

AFLP ANALYSIS OF ARTIFICIAL GYNOGENESIS IN PSEUDOSCIAENA CROCEA

A comparative analysis of AFLP was performed by using 6 pairs of selective primers (E-AAG/M-CAG, E-AAG/M-CTC, E-ACA/M-CAA; E-ACG/M-CAT, E-AGG/M-CAA, E-AGG/M-CTA) in Pseudociaena crocea fries from two groups(G1, G2)of the gynogens induced by cool-shock method, two groups of contrast (C1, C2)and their parents. The similarity coefficient and genetic distance among gynogenetic offspring, the contrast(hybrid)and their parents were calculated along with phylogenetic clustering analysis.A total of 33 special male bands(13 in G1, 20 in G2),and 31 special female bands(16 in G1, 15 in G2)were revealed in 6 pairs of primers.The amplification showed no male gene in 24 fries of family G1, suggesting that they all derived from gynogenesis.The successful induction ratio was 100%.Different special male bands appeared in 3 individuals of family G2, suggesting the origin of normal fertilization(12.5%).Its successful induction ratio was 87.5%.The average gynogenetic ratio of the two families was 93.75%.Thirteen maternal loci(8 in G1, 5 in G2)were homozygotic. 14 of 31 special female bands were separated in the gynogens indicative of heterozygosity. The homozygosity ratio of the offspring from the two families was 87.5% and 76.2% in average of 81.9%, whereas it was zero in their contrast. It is clear that the similarity coefficient among gynogens of two families was the largest, or the genetic distance among gynogens was the shortest among the hybrids of their contrast.The relation of the gynogens was the closest to the female parent but male one.Therefore, gynogenesis analysis with AFLP marker technique is practical in gene purification, verification and other genetic analyses.